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作 者:袁茵[1] 王一飞[1] 张美英[1] 熊盛[1] 李久香[1] 胡红梅 刘秋英[1] 吴志聪[1]
出 处:《暨南大学学报(自然科学与医学版)》2004年第4期400-407,共8页Journal of Jinan University(Natural Science & Medicine Edition)
基 金:国家自然科学基金资助项目 (30 3716 6 1) ;国家十五重点攻关"86 3"项目 (2 0 0 1AA2 15 0 4 1)
摘 要:目的 :探讨nm2 3-H1基因的表达与白血病细胞HL - 6 0增殖之间的关系。方法 :以 2 5ng/mL阿糖胞苷处理HL - 6 0细胞 ,MTT法测定细胞生长抑制率 ,NBT还原比色法判断细胞分化状况 ,RT -PCR检测nm2 3-H1基因表达的变化 ;构建nm2 3-H1基因的真核表达质粒pEGFP -N1-nm2 3-H1,转染HL - 6 0细胞 ,通过细胞生长曲线和血清依赖性实验检测nm2 3-H1基因的过表达对HL - 6 0细胞生长的影响。结果 :小剂量Ara -C对HL - 6 0细胞的生长呈时间依赖性抑制 ,作用4d后细胞NBT还原能力增强且nm2 3-H1基因的表达下调 ;转染nm2 3-H1基因的HL - 6 0细胞生长加快、血清依赖性下降。结论 :Ara -C对HL - 6 0细胞增殖的抑制作用与下调nm2 3-H1基因的表达有一定关系 ;nm2 3-H1基因在HL - 6 0细胞中的过表达有促细胞增殖的作用 ,即增高了HL -6To investigate the relationship between nm23-H1 gene expression and proliferation of HL-60 cells. Methods: The changes of proliferation rate, differentiation and nm23-H1 gene expression in HL-60 cells treated with low-dose cytosine arabinoside were observed by MTT assay, NBT reduction assay and RT-PCR. The recombined eukaryotic expression vector pEGFP-N1-nm23-H1 was constructed to transfect HL-60 cells. The effect of nm23-H1 gene overexpression on HL-60 cells was analyzed through cell growth curve and serum-dependent test. Results: Low-dose cytosine arabinoside could suppress the proliferation of HL-60 cells in a time-dependent way and lower the expression of nm23-H1 gene. 42 percent of the HL-60 cells obtained the NBT reduction ability after being treated with 25 ng/mL Ara-C for 4 d. Overexpression of nm23-H1 stimulated the growth of HL-60 cells and reduced the dependence on serum of them. Conclusion: The inhibitory effect of Ara-C on HL-60 cell proliferation had a certain relationship with the down-regulation of nm23-H1 gene, and the overexpression of nm23-H1 gene in HL-60 cells improved cell proliferation, namely elevated malignant degree of them. nm23-H1 gene might be a potential target of leukemia treatment.
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